鉴定单增李斯特菌4b血清型亚型的多重PCR方法的建立

Development of a multiplex PCR assay for differentiation of Listeria monocytogenes serotype 4b subgroups

目的利用多重PCR(multiplex PCR)技术,建立针对单增李斯特菌4b血清型亚型(ECI、ECII、ECIV、non-EC)的鉴定方法。方法以LMOf2365_2798、LMOh7858_0487、ORF2110和gtcA作为靶基因设计引物,建立同时鉴别ECI、ECII、ECIV、non-EC的多重PCR反应体系,并对16株4b血清型单增李斯特菌进行亚型鉴定。结果 16株4b血清型单增李斯特菌可用本方法进行分型:其中ECI亚型11株,ECII亚型1株,ECIV亚型2株,非流行克隆群(non-EC)2株。结论本方法能准确鉴定单增李斯特菌4b血清型菌株的4个亚型,可用于食品、临床标本的病原学诊断和疫情暴发的溯源调查。

A novel multiplex PeR assay was developed to rapidly separate L. monocytogenes serotype 4b strains into four subgroups （ECI, ECII, ECIV, non-EC） by simultaneously targeting LMOf2365_2798, LMOh78580487, ORF2110, and gtcA. The PCR test was successfully evaluated with 16 serotype 4b isolates, with 11 isolates belonging to ECI, one belonging to ECII, two belonging to ECⅢ , and two belonging to non-EC. The novel one-step molecular method will be valuable and reli able for rapid discrimination of the distinct subgroups strains of L. monocytogenes serotype 4b from food samples, processing environments and clinical samples, which would facilitate the source tracing of the outbreak of L. monocytogenes in the future.